Figure 4

A. DPV UL46 and UL46M gene encoding DNA sequences were cloned into pET32a (+) procaryotic expression vector. The recombinant plasmids were digested with two restriction enzymes. Lane 1, Bam HI and Xho I generating two restriction fragments of UL46; Lane M, DNA marker; Lane 2, Bam HI and Xho I generating two restriction fragments of UL46M. B. Induction of the 6× His-tagged UL46M fusion protein in E. coli plasmid pET32-a (+)/UL46M was transformed into bacteria. Bacteria were grown in the absence (lane 4) or the presence (lane2 and lane 3) of IPTG. Induced pET32-a (+) was as control (lane 1). Molecular mass marker (in kDa) were shown to the right (lane 5). C. The recombinant UL46M fusion protein was purified by Ni-NTA affinity chromatography. Lane 1, unpurified recombinant UL46M fusion protein; Lane 2, purified recombinant UL46M fusion protein; Lane M, protein marker. D. Detection result of the antisera titer by agar diffusion reaction. The result of the agar diffusion reaction of the anti-UL46M antiserum showed the largest dilution multiple of the positivation was 1:8. 1-6.1, 2, 4, 8, 16, 32-fold diluted antisera; 0. DPV. E. Analysis of the antibody specificity by western blot The result revealed that the purified recombinant protein was recognized by the anti-UL46M rabbit IgG and showed a specific signal at the expected size (79 kDa). No positive signal was observed when using the negative control sera (date not shown). Lane 1, western blot of anti-DPV IgG with the UL46M protein; Lane 2, western blot of anti-DPV UL46M IgG with the UL46M protein; Lane M, prestained protein marker.